Figure 1.

Schematic representation of the workflow to perform lysine acetylation stoichiometric analysis following the three sample preparation procedures, FASP, GSP, and SSP. In all strategies, untargeted lysine residues are chemically acetylated with the deuterated derivate of NHS (NAS-d₃), and side reactions are reverted by treating proteins with hydroxylamine. To avoid unwanted protein precipitation in FASP and SSP, trypsin digestion was performed in the presence of SDC. Peptide mixtures were analyzed in a high-resolution LC-MS/MS system. Data analysis was performed by means of MaxQuant and Pview software.